Characterization of cDNA clones for the alpha subunit of pyruvate dehydrogenase from Ascaris suum

Johnson, Keith R.; Komuniecki, Richard; Sun, Yinghao; Wheelock, Margaret J.

doi:10.1016/0166-6851(92)90198-s

Cited by 26 publications

(8 citation statements)

References 38 publications

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“…The highest score (38% identity) was obtained with the deduced primary structure of the S. avermitilis bkdA gene (E1␣ subunit) (39). Lower sequence identities (Ͻ29%) to the E1␣ subunits of other PDH complexes were detected (4,22,24). A multiple sequence alignment identified common structural motifs well conserved in the E1␣ subunits of all of the ␣-keto acid dehydrogenase complexes examined so far, such as FIG.…”

Section: Resultsmentioning

confidence: 98%

“…Table 1 (48); B. subtilis (19) and B. stearothermophilus (18) E1␣ dual PDH-BCDHs (PB-Bsu and PB-Bst, respectively); A. laidlawii partially characterized E1␣ PDH (P?-Ala) (47); bovine (21) and human (16) E1␣ BCDHs (B-Bta and B-Hsa, respectively); and Saccharomyces cerevisiae (4), Ascaris suum (22), human (24), and rat (10a) E1␣ PDHs (P-Sce, P-Asu, P-Hsa, and P-Rra, respectively). Conserved positions are indicated as follows: boldface, residues identical to those in the S. avermitilis bkdF gene product; F, residues identical in all sequences; E, residues either identical or conservatively substituted in the S. exhibited a typical bkd mutant phenotype (the other three thiostrepton-sensitive, erythromycin-resistant clones mentioned above also had a similar phenotype).…”

Section: Vol 177 1995 S Avermitilis Bkdf Mutant and Avermectin Promentioning

confidence: 99%

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A second branched-chain alpha-keto acid dehydrogenase gene cluster (bkdFGH) from Streptomyces avermitilis: its relationship to avermectin biosynthesis and the construction of a bkdF mutant suitable for the production of novel antiparasitic avermectins

et al. 1995

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A second cluster of genes encoding the E1␣, E1␤, and E2 subunits of branched-chain ␣-keto acid dehydrogenase (BCDH), bkdFGH, has been cloned and characterized from Streptomyces avermitilis, the soil microorganism which produces anthelmintic avermectins. Open reading frame 1 (ORF1) (bkdF, encoding E1␣), would encode a polypeptide of 44,394 Da (406 amino acids). The putative start codon of the incompletely sequenced ORF2 (bkdG, encoding E1␤) is located 83 bp downstream from the end of ORF1. The deduced amino acid sequence of bkdF resembled the corresponding E1␣ subunit of several prokaryotic and eukaryotic BCDH complexes. An S. avermitilis bkd mutant constructed by deletion of a genomic region comprising the 5 end of bkdF is also described. The mutant exhibited a typical Bkd ؊ phenotype: it lacked E1 BCDH activity and had lost the ability to grow on solid minimal medium containing isoleucine, leucine, and valine as sole carbon sources. Since BCDH provides an ␣-branched-chain fatty acid starter unit, either S(؉)-␣-methylbutyryl coenzyme A or isobutyryl coenzyme A, which is essential to initiate the synthesis of the avermectin polyketide backbone in S. avermitilis, the disrupted mutant cannot make the natural avermectins in a medium lacking both S(؉)-␣-methylbutyrate and isobutyrate. Supplementation with either one of these compounds restores production of the corresponding natural avermectins, while supplementation of the medium with alternative fatty acids results in the formation of novel avermectins. These results verify that the BCDH-catalyzed reaction of branched-chain amino acid catabolism constitutes a crucial step to provide fatty acid precursors for antibiotic biosynthesis in S. avermitilis.

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Section: Resultsmentioning

confidence: 98%

Section: Vol 177 1995 S Avermitilis Bkdf Mutant and Avermectin Promentioning

confidence: 99%

A second branched-chain alpha-keto acid dehydrogenase gene cluster (bkdFGH) from Streptomyces avermitilis: its relationship to avermectin biosynthesis and the construction of a bkdF mutant suitable for the production of novel antiparasitic avermectins

et al. 1995

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“…A comparison of the percentage identities (second number) and percentage similarities (first number) obtained when the predicted amino acid sequences of Ela subunits from different sources are compared using the GAP program (Devereux et a/., 1984) cerevisiae (Behal et al, 1989), A. suum (Johnson et al, 1992), mouse (Fitzgerald et al, 19921, and human (Dahl et al, 1990).…”

Section: E1 a Gene Expressionmentioning

confidence: 99%

Mitochondrial Pyruvate Dehydrogenase (Molecular Cloning of the E1[alpha] Subunit and Expression Analysis)

et al. 1995

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The mtPDC occupies a central point in cellular energy metabolism, linking glycolytic carbon metabolism with the citric acid cycle. Since the reaction catalyzed by mtPDC is irreversible, this represents the committed step of carbon entry into the citric acid cycle. Plants are unique in that, in addition to mtPDC, they contain a second isoform of PDC localized in plastids. This isoform is believed to be involved in the provision of acetyl-COA for fatty acid, isoprenoid, and amino acid biosynthesis (Camp and Randall, 1985).Our knowledge of the structural organization of mtPDC is based largely on studies of the mammalian enzyme. The complex has a molecular weight of approximately 7 x 106 and is composed of enzymic, regulatory, and structural components. It contains the enzymes E1 (EC 1.2.4.1), E2 '

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“…FASTA analysis showed the protein product to be the a-subunit of pyruvate dehydrogenase Enzyme I. Figure 5 shows (Ho et al ., 1989), and (odpt-human) (Dah1 et al, 1987); pig (odpa-pig) (Sermon et al, 1990); mouse (odpa-mouse) (Fitzgerald et al, 1992) and (odpt-mouse) (Fitzgerald et al, 1992); rat (odba-rat) (Zhang et al, 1987), (odpt-rat) (Cullingford et a]., 1993), and (odpa-rat) (Cullingford et al, 1994); Ascarus suum (odpt-ascu) (Johnson et al, 1992) and (odpa-ascu) (Johnson et al, 1992); C. magnum (acoa-cloma) (Kruger et a]., 1994); A . eutrophus (acoaalceu) (Priefert et al, 1991); Pelobucter carbinolicus (acoa-pelca) ; K. pneumoniae (acoa-klepn) (Deng et al, 1994); bovine (odba-bovin) ; Pseudomonasputidu (odba-psepu) (Burns et al, 1988).…”

Section: O O a I L C " T P A T T A O A T T M T~t A T~~a~m C -~A~~g K~unclassified

Sequence and organization of genes encoding enzymes involved in pyruvate metabolism in mycoplasma capricolum

Zhu

Peterkofsky

1996

Protein Science

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The region of the genome of Mycoplasma capricolum upstream of the portion encompassing the genes for Enzymes I and IIAg" of the phosphoeno1pyruvate:sugar phosphotransferase system (PTS) was cloned and sequenced. Examination of the sequence revealed open reading frames corresponding to numerous genes involved with the oxidation of pyruvate. The deduced gene organization is naox (encoding NADH oxidase)-lplA (encoding lipoate-protein ligase)-odpA (encoding pyruvate dehydrogenase EIa)-odpB (encoding pyruvate dehydrogenase EIP)-odp2 (encoding pyruvate dehydrogenase EI1)-dldH (encoding dihydrolipoamide dehydrogenase)-pta (encoding phosphotransacety1ase)-ack (encoding acetate kinase)-orfA (an unknown open reading frame)-kdtB-ptsl-crr. Analysis of the DNA sequence suggests that the naox and IpIA genes are part of a single operon, odpA and odpB constitute an additional operon, odp2 and dldH a third operon, andpta and ack an additional transcription unit. Phylogenetic analyses of the protein products of the odpA and odpB genes indicate that they are most similar to the corresponding proteins from Mycoplasma genitalium, Acholeplasma laidlawii, and Gram-positive organisms. The product of the odp2 gene contains a single lipoyl domain, as is the case with the corresponding proteins from M. genitalium and numerous other organisms. An evolutionary tree places the M. capricolum odp2 gene product in close relationship to the corresponding proteins from A. laidlawii and M. genitalium. The dldH gene encodes an unusual form of dihydrolipoamide dehydrogenase that contains an aminoterminal extension corresponding to a lipoyl domain, a property shared by the corresponding proteins from Alcaligenes eutrophus and Clostridium magnum. Aside from that feature, the protein is related phylogenetically to the corresponding proteins from A. laidlawii and M. genitalium. The phosphotransacetylase from M. capricolum is related most closely to the corresponding protein from M. genitalium and is distinguished easily from the enzymes from Escherichia coli and Haemophilus influenzae by the absence of the characteristic amino-terminal extension. The acetate kinase from M. capricolum is related evolutionarily to the homologous enzyme from M. genitalium. Map position comparisons of genes encoding proteins involved with pyruvate metabolism show that, whereas all the genes are clustered in M. capricolum, they are scattered in M. genitalium.

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Characterization of cDNA clones for the alpha subunit of pyruvate dehydrogenase from Ascaris suum

Cited by 26 publications

References 38 publications

A second branched-chain alpha-keto acid dehydrogenase gene cluster (bkdFGH) from Streptomyces avermitilis: its relationship to avermectin biosynthesis and the construction of a bkdF mutant suitable for the production of novel antiparasitic avermectins

A second branched-chain alpha-keto acid dehydrogenase gene cluster (bkdFGH) from Streptomyces avermitilis: its relationship to avermectin biosynthesis and the construction of a bkdF mutant suitable for the production of novel antiparasitic avermectins

Mitochondrial Pyruvate Dehydrogenase (Molecular Cloning of the E1[alpha] Subunit and Expression Analysis)

Sequence and organization of genes encoding enzymes involved in pyruvate metabolism in mycoplasma capricolum

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